Di Pasquale Elisa, Song Belle, Condorelli Gianluigi
Humanitas Clinical and Research Center, Italy.
J Vis Exp. 2013 Jun 28(76):50429. doi: 10.3791/50429.
In order to investigate the events driving heart development and to determine the molecular mechanisms leading to myocardial diseases in humans, it is essential first to generate functional human cardiomyocytes (CMs). The use of these cells in drug discovery and toxicology studies would also be highly beneficial, allowing new pharmacological molecules for the treatment of cardiac disorders to be validated pre-clinically on cells of human origin. Of the possible sources of CMs, induced pluripotent stem (iPS) cells are among the most promising, as they can be derived directly from readily accessible patient tissue and possess an intrinsic capacity to give rise to all cell types of the body (1). Several methods have been proposed for differentiating iPS cells into CMs, ranging from the classical embryoid bodies (EBs) aggregation approach to chemically defined protocols (2,3). In this article we propose an EBs-based protocol and show how this method can be employed to efficiently generate functional CM-like cells from feeder-free iPS cells.
为了研究驱动心脏发育的事件并确定导致人类心肌疾病的分子机制,首先必须生成功能性人类心肌细胞(CMs)。在药物发现和毒理学研究中使用这些细胞也将非常有益,这使得用于治疗心脏疾病的新药理学分子能够在源自人类的细胞上进行临床前验证。在CMs的可能来源中,诱导多能干细胞(iPS细胞)是最有前景的来源之一,因为它们可以直接从容易获取的患者组织中获得,并且具有产生身体所有细胞类型的内在能力(1)。已经提出了几种将iPS细胞分化为CMs的方法,从经典的胚状体(EBs)聚集方法到化学成分明确的方案(2,3)。在本文中,我们提出了一种基于EBs的方案,并展示了如何使用该方法从无饲养层的iPS细胞中高效生成功能性类CM细胞。